title: Quantifying elongation rhythm during full-length protein synthesis.
abstract: Pauses regulate the rhythm of ribosomal protein synthesis. Mutations disrupting even minor pauses can give rise to improperly formed proteins and human disease. Such minor pauses are difficult to characterize by ensemble methods, but can be readily examined by single-molecule (sm) approaches. Here we use smFRET to carry out real-time monitoring of the expression of a full-length protein, the green fluorescent protein variant Emerald GFP. We demonstrate significant correlations between measured elongation rates and codon and isoacceptor tRNA usage, and provide a quantitative estimate of the effect on elongation rate of replacing a codon recognizing an abundant tRNA with a synonymous codon cognate to a rarer tRNA. Our results suggest that tRNA selection plays an important general role in modulating the rates and rhythms of protein synthesis, potentially influencing simultaneous co-translational processes such as folding and chemical modification.
authors.affiliation: Department of Chemistry, University of Pennsylvania, Philadelphia, Pennsylvania 19104-6323, USA.
authors.forename: Gabriel
authors.initials: G
authors.lastname: Rosenblum
authors.forename: Chunlai
authors.initials: C
authors.lastname: Chen
authors.forename: Jaskiran
authors.initials: J
authors.lastname: Kaur
authors.forename: Xiaonan
authors.initials: X
authors.lastname: Cui
authors.forename: Haibo
authors.initials: H
authors.lastname: Zhang
authors.forename: Haruichi
authors.initials: H
authors.lastname: Asahara
authors.forename: Shaorong
authors.initials: S
authors.lastname: Chong
authors.forename: Zeev
authors.initials: Z
authors.lastname: Smilansky
authors.forename: Yale E
authors.initials: YE
authors.lastname: Goldman
authors.forename: Barry S
authors.initials: BS
authors.lastname: Cooperman
mesh_terms: D000595:Amino Acid Sequence; D003062:Codon / Q000235:genetics; D031541:Fluorescence Resonance Energy Transfer; D049452:Green Fluorescent Proteins / Q000737:chemistry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D010441:Peptide Chain Elongation, Translational; D010446:Peptide Fragments / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D011487:Protein Conformation; D012343:RNA, Transfer / Q000235:genetics; D012270:Ribosomes / Q000235:genetics / Q000378:metabolism
chemical_list: D003062:Codon; D010446:Peptide Fragments; D049452:Green Fluorescent Proteins; D012343:RNA, Transfer